High Levels of Biologically Active Vascular Endothelial Growth Factor (VEGF) are Produced by the Baculovirus Expression System

1992 ◽  
Vol 7 (2) ◽  
pp. 131-138 ◽  
Author(s):  
Tzafra Cohen ◽  
Hela Gitay-Goren ◽  
Gera Neufeld ◽  
Ben-Zion Levi
2020 ◽  
Vol 24 (4) ◽  
pp. 114
Author(s):  
V. V. Sevostianova ◽  
A. V. Mironov ◽  
L. V. Antonova ◽  
E. O. Krivkina ◽  
V. G. Matveeva ◽  
...  

<p><strong>Background.</strong> Commercially available synthetic and animal-derived vascular patches used in patch angioplasty during carotid endarterectomy have several disadvantages, such as postoperative thrombosis or occlusion and restenosis. This problem may be resolved by the development of biologically active materials that are biodegradable and can stimulate tissue regeneration.<br />Aim. To evaluate the properties and efficacy of a biodegradable patch based on poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL) into which vascular endothelial growth factor (VEGF) is incorporated, in comparison with unmodified PHBV/PCL and commercial vascular patches.</p><p><strong>Methods.</strong> Porous patches were fabricated by emulsion electrospinning from a mixture of PHBV and PCL, into which VEGF was incorporated. The morphological and mechanical properties of these patches were tested, and they were implanted into the wall of rat abdominal aortas for 1, 3, 6 and 12 months. Histological and immunofluorescence examinations were performed to evaluate endothelisation, cellular composition and calcification.</p><p><strong>Results.</strong> PHBV / PCL patches with VEGF had a highly porous structure and demonstrated tensile strength similar to that of the aorta in rats and the internal thoracic artery in humans. After 3 months of implantation, an endothelial monolayer was formed on the inner surface of these patches. The patches were populated by cells that secreted the extracellular matrix faster than did cells of patches from the xenopericardium. Remodelling with PHBV / PCL patches was not accompanied by chronic inflammation; in contrast, inflammation was observed with long-term implantation of unmodified PHBV / PCL samples.</p><p><strong>Conclusion.</strong> VEGF incorporated into biodegradable PHBV / PCL patches stimulated their endothelisation, increased their biocompatibility and promoted remodelling and formation of the components of the blood vessel. PHBV / PCL / VEGF patches thus have a high potential for use in tissue engineering of the vascular wall.</p><p>Received 2 June 2020. Revised 27 June 2020. Accepted 16 July 2020.</p><p><strong>Funding:</strong> This study was supported by the Complex Program of Basic Research under the Siberian Branch of the Russian Academy of Sciences within the Basic Research Topic of Research Institute for Complex Issues of Cardiovascular Diseases № 0546-2019-0002 “Pathogenetic basis for the development of cardiovascular implants from biocompatible materials using patient-oriented approach, mathematical modeling, tissue engineering, and genomic predictors”.</p><p><strong>Conflict of interest:</strong> Authors declare no conflict of interest.</p><p><strong>Author contributions</strong><br />Conception and study design: V.V. Sevostianova, A.V. Mironov, L.V. Antonova, R.S. Tarasov, L.S. Barbarash<br />Data collection and analysis: V.V. Sevostianova, A.V. Mironov, L.V. Antonova, E.O. Krivkina, V.G. Matveeva, E.A. Velikanova, T.V. Glushkova<br />Statistical analysis: V.V. Sevostianova, T.V. Glushkova<br />Drafting the article: V.V. Sevostianova, A.V. Mironov <br />Critical revision of the article: L.V. Antonova, R.S. Tarasov, L.S. Barbarash<br />Final approval of the version to be published: V.V. Sevostianova, A.V. Mironov, L.V. Antonova, E.O. Krivkina, V.G. Matveeva, E.A. Velikanova, R.S. Tarasov, T.V. Glushkova, L.S. Barbarash</p>


2011 ◽  
Vol 164 (1) ◽  
pp. 115-122 ◽  
Author(s):  
Anniina Färkkilä ◽  
Mikko Anttonen ◽  
Jurate Pociuviene ◽  
Arto Leminen ◽  
Ralf Butzow ◽  
...  

ObjectiveOvarian granulosa cell tumors (GCTs) are hormonally active sex cord stromal tumors accounting for 3–5% of all ovarian cancers. These tumors are generally diagnosed at an early stage but there is a high risk of recurrence, associated with high mortality. Treatment of recurrent GCTs is difficult, and biologically targeted treatment modalities are lacking. GCTs are highly vascularized, and angiogenic factors most probably play a role in their pathology. Vascular endothelial growth factor (VEGF) is a key regulator of tumor angiogenesis, but in GCTs, the role of VEGF and its receptors VEGFR-1 (FLT1) and VEGFR-2 (KDR) remains largely unknown. Our objective is to study the expression of VEGF and its receptors in human GCTs.MethodsWe analyzed GCTs from 106 patients for the expressions of VEGF and its receptors utilizing tumor tissue microarray, tumor mRNA, and patient serum samples.ResultsWe found that VEGF and its main biologically active receptor VEGFR-2 were highly expressed in primary and recurrent GCTs, when compared with normal granulosa-lutein cells. The expression of VEGF correlated positively to tumor microvessel density and to VEGFR-2 expression at the protein (P<0.05) and mRNA (P<0.05) levels. In contrast to VEGFR-2, the expression of VEGFR-1 was weak. Tumor VEGF protein expression was not prognostic for recurrence, however, we found high levels of circulating VEGF in the serum of patients with primary GCT.ConclusionsThe results suggest an important role of VEGF and VEGFR-2 in GCT pathology and support the possibility of applying novel VEGF- or VEGFR-2-targeted treatments to patients with GCT.


2020 ◽  
Author(s):  
Hülya Kuduğ Ceylan ◽  
Seçil Erden Tayhan ◽  
İsa Gökçe

Abstract Among the vascular endothelial growth factor (VEGF) family variants, the 165-amino acid isoform (VEGF165) is the best characterized and most potent endothelial cell mitogenic factor. It is known that VEGF165 mediates angiogenesis and has the potential for the therapeutic applications. In this study, the expression system of Kluyveromyces lactis that produces the recombinant human VEGF165 has been evaluated. The gene encoding human VEGF165 was successfully cloned in the pKLAC2 expression vector containing a strong LAC4 promoter, after which a pKLAC2-VEGF165 plasmid was constructed. After the transformation, the recombinant human vascular endothelial growth factor 165 (rhVEGF165) was expressed in K. lactis GG799 cells (~ 5.7 mg/L) confirmed by SDS-PAGE and Western blotting and downstream purification processed comprising ammonium sulphate precipitation and affinity chromatography. The biological activity of the purified rhVEGF165 was confirmed by the proliferation of the human umbilical vein-derived endothelial cells (HUVEC) in a dose- and time-dependent manner. The K. lactis-derived rhVEGF165 exhibited a higher proliferative activity compared with a commercially available rhVEGF165 with a half-maximal effective concentration of 3.0. Cell migration analysis was conducted to evaluate the in vitro wound healing effect of the produced rhVEGF165 via a scratch assay. These findings indicate that K. lactis could be a suitable host for secreting bioactive human VEGF165 for therapeutic use.


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